Nonviral production of human interleukin-7 in spodoptera frugiperda insect cells as a soluble recombinant protein.
Human interleukin-7 (hIL-7) is a cytokine secreted by the stromal cells of the red marrow. It is important for proliferation during certain stages of B-cell maturation and for T and NK cell survival, development, and homeostasis. It is a critical growth factor for enhancement and recovery of the immune T-cell. Because of its strong immunomodulatory effects, hIL-7 may become a valuable supplementary agent for immunotherapeutical treatments in patients with HIV infection or immunodeficiency. Human IL-7 has previously been produced in various protein expression systems. In this paper, we present an alternative expression system, in Spodoptera frugiperda cells, for the production of hIL-7 using nonlytic vector systems. This system allows generation of correctly translated and accurately processed heterologous proteins as soluble recombinant proteins. Here we report plasmid construction, transfection, and consequent expression of hIL-7 using this nonlytic insect cell expression system. The levels of secreted hIL-7 in a small scale experiment reached a level of 1.7 microg x 1(-1) under serum-free cell culture conditions.
Mirzaei
Maryam
M
Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering, and Artificial Cells and Organs Research Centre, Faculty of Medicine, McGill University, 3775 University Street, Montreal, PQ, Canada H3A 2B4.
Xu
Yan
Y
Elias
Cynthia B
CB
Prakash
Satya
S
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D012831:Bombyx / Q000235:genetics; D002461:Cell Line, Transformed; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D015851:Interleukin-7 / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D015202:Protein Engineering / Q000379:methods; D011691:Puromycin; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D018411:Spodoptera / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D015851:Interleukin-7; D011993:Recombinant Fusion Proteins; D011691:Puromycin